Zinc Finger Consortium: Zinc Finger Arrays

Engineered Zinc Finger Arrays for Endogenous Zebrafish Gene Targets

Recent published reports have shown that engineered zinc finger nucleases (ZFNs) are effective tools for making targeted mutations in zebrafish. With support from the NIH and guidance from the academic community, the labs of Keith Joung, Randall Peterson, and Joanna Yeh are engineering zinc finger arrays targeted to various zebrafish genes. The table below lists zebrafish genes and associated pairs of zinc finger arrays that can be used to generate ZFNs targeted to a site within that gene.

To construct ZFN expression vectors, DNA encoding zinc finger arrays should be cloned into plasmids also available through Addgene--the specific vector to use depends on the length of the spacer sequence between the half-sites (see Nuclease Expression Vectors for more details).

IMPORTANT NOTE: All zinc finger arrays listed show transcriptional activation activity of three-fold or greater in a well-characterized bacterial two-hybrid reporter system but have NOT yet been tested for activity as ZFNs in zebrafish.

Scientists using these zinc finger arrays in a publication should notify [email protected] and acknowledge NIH grant number R01 GM088040 in the publication.

References

• Rapid 'open-source' engineering of customized zinc-finger nucleases for highly efficient gene modification. Maeder ML et al. Mol Cell.. 2008. Jul 25. 31(2):294-301. PubMed PMID 18657511.

• Oligomerized pool engineering (OPEN): an 'open-source' protocol for making customized zinc-finger arrays. Maeder ML et al. Nat Protoc.. 2009. Sept 17. 4(10):1471-1501. PubMed PMID 19798082.

• Targeted mutagenesis in zebrafish using customized zinc-finger nucleases. Foley JE et al. Nat Protoc.. 2009. Dec 3. 4(12):1855-1867. PubMed PMID 20010934.